Abstract
Alkaline proteinase produced by one of the monospore isolates of Streptomyces violaceorectus MC675-A8, a producer of the alkaline metalloendopeptidases named alkinonases A and AF, was purified by affinity chromatography on N-benzyloxycarbonylglycylleucylaminohexyl-Sepharose to electrophoretic homogeneity. The enzyme was inactivated by phenylmethanesulfonyl fluoride and diisopropylfluorophosphate, but not by chelating agents or sulfhydryl reagents, and was distinct from alkinonases A and AF. The optimum pH for casein hydrolysis was 9.5-10.5, and the enzyme was stable within a pH range of 5.0-12.0. The molecular weight was estimated to be 30000. The peptidase activity of the enzyme was greatest on N-benzyloxycarbonylglycylprolylleucylglycine ethyl ester among the synthetic substrates tested in this work. Amidase activity was observed towards peptide amides such as N-benzyloxycarbonylglycylleucine amide and N-benzyloxycarbonylglycylphenylalanine amide. The enzyme showed anti-inflammatory activity against carrageenan-induced edema in rats, as did alkinonases A and AF. Bradykinin was hydrolyzed by the enzyme to arginylprolylprolylglycylphenylalanylserylprolylphenylalanine and arginine. Since the enzyme showed a serine proteinase nature, the enzyme was named alkaline proteinase S, and the producing microbe was designated as Streptomyces violaceorectus MC675-A8-S.
